Additional file 2 of Gut microbiota-testis axis: FMT improves systemic and testicular micro-environment to increase semen quality in type 1 diabetes

Additional file 2: Fig. S1. Body weight and gut microbiota changes (STZ vs. Con). a Animal bodyweight. The y-axis represents the body weight (g). The x-axis represents the age (weeks). b Blood insulin levels. The y-axis represents the concentration (mIU/L). The x-axis represents the treatment. c The alpha index of the small intestine microbiota (Chao index). The y-axis represents the relative amount. The x-axis represents the treatment. d The beta index of small intestinal microbiota. The y-axis represents the relative amount. The x-axis represents the treatment. e PLS-DA (OTU) of small intestine microbiota in STZ and Con groups. f Small intestine microbiota levels at the genus level in STZ and Con groups. The y-axis represents the relative amount (%). The x-axis represents the individual microbiota. g The alpha index of the cecum microbiota (Chao index). The y-axis represents the relative amount. The x-axis represents the treatment. h The beta index of cecum microbiota. The y-axis represents the relative amount. The x-axis represents the treatment. i PLS-DA (OTU) of cecum microbiota in STZ and Con groups. j Cecum microbiota levels at the genus level in STZ and Con groups. The y-axis represents the relative amount (%). The x-axis represents the individual microbiota. k The alpha index of the colon microbiota (Chao index). The y-axis represents the relative amount. The x-axis represents the treatment. l The beta index of colon microbiota. The y-axis represents the relative amount. The x-axis represents the treatment. m PLS-DA (OTU) of colon microbiota in STZ and Con groups. n Colon microbiota levels at the genus level in STZ and Con groups. The y-axis represents the relative amount (%). The x-axis represents the individual microbiota. Fig. S2. a PLS-DA (OTU) of cecum microbiota in STZ, A10-FMT, and Con-FMT groups. b Cecum microbiota levels at the genus level in STZ, A10-FMT, and Con-FMT groups. The y-axis represents the relative amount (%). The x-axis represents the treatments. Different colors represent different microbiota. c Cladogram of the LEfSe determining the cecum microbiota difference in abundance. d Changed microbiota in the cecum. The y-axis represents the relative amount at the genus level. The x-axis represents the treatment. *p < 0.05. e PLS-DA (OTU) of colon microbiota in STZ, A10-FMT, and Con-FMT groups. f Colon microbiota levels at the genus level in STZ, A10-FMT, and Con-FMT groups. The y-axis represents the relative amount (%). The x-axis represents the treatments. Different colors represent different microbiota. g Cladogram of the LEfSe determining the difference in abundance of colon microbiota. h Changed microbiota in the colon. The y-axis represents the relative amount at the genus level. The x-axis represents the treatment. *p < 0.05. Fig. S3. Blood metabolism changes. a Blood flavonoid Malvidin 3-O-(6-O-(4-O-feruloyl-alpha-rhamnopyranosyl)-beta-glucopyranoside)-5-beta-glucopy -ranoside levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. b Blood glycerophosphocholine levels in different treatments. The y-axis represents the fold change compared to control group (Con). The x-axis represents the treatment. c Blood LysoPC (15:0) levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. d Blood LysoPC [18:1(9z)] levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. e Blood LysoPC [22:6(4Z,7Z,10Z,13Z,16Z,19Z)] levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. f Blood PE [22:2(13Z,16Z)/0:0] levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. g Blood PE (22:0/0:0) levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. h Blood PE [22:1(11Z)/0:0] levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. i Blood LysoPE [0:0/24:6(6Z,9Z,12Z,15Z,18Z,21Z)] levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. Fig. S4. Testicular metabolite changes. a Testicular 4,8,12,15,18-eicosapentaenoic acid levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. b Testicular EPA in different treatments. The y-axis represents the relative amount. The x-axis represents the treatment. *p < 0.05. c Testicular retinoids levels in different treatments. The y-axis represents the fold change compared to the control group (Con). The x-axis represents the treatment. d Testicular retinol levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. e Testicular 9-cis-retinal levels in different treatments. The y-axis represents the relative amount. The x-axis represents the treatment. *p < 0.05. f Testicular retinyl ester levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. g Quantitative data for Western blotting analysis in Fig. 4d. h Testicular testosterone levels in different treatments. The y-axis represents the fold change compared to control group (Con). The x-axis represents the treatment. i Testicular testosterone propionate levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. j Testicular testosterone acetate levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. k Testicular 7alpha-hydroxytestosterone levels in different treatments. The y-axis represents the relative amount. The x-axis represents the treatment. *p < 0.05. l Testicular steroids levels in different treatments. The y-axis represents the fold change compared to control group (Con). The x-axis represents the treatment. m Testicular 3b,17b-Dihydroxyetiocholane levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. n Testicular 5-Androstene-3b,16b,17a-triol levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. o Testicular 3beta,17alpha,21-Trihydroxy-pregnenone levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. p Testicular asterogenol levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. q Testicular PCs levels in different treatments. The y-axis represents the fold change compared to the control group (Con). The x-axis represents the treatment. r Testicular) PC(4:0/18:1(9Z) level. The y-axis represents the relative amount (%). The x-axis represents the treatments. s Testicular PC (8:0/8:0) levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. t Testicular 6-hydroxymelatonin levels in different treatments. The y-axis represents the relative amount. The x-axis represents the treatment. *p < 0.05. Fig. S5. Additional data for testis and spleen. a Western blotting analysis of the proteins of some important genes for spermatogenesis in each treatment. n≥3/group. b PCA for RNA-seq analysis of spleen. c The functional enrichment analysis of STZ increased genes while these were decreased by A10-FMT or Con-FMT. d IHF staining of ki67 in the spleen. e Western blotting analysis of p53, Bax and Bcl-xl in the spleen. Fig. S6. A10-FMT improved liver function and systemic anti-oxidative capability. a Blood alanine aminotransferase (ALT) levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. b PCA for RNA-seq analysis of liver. c The functional enrichment analysis of STZ decreased genes while these were increased by A10-FMT or Con-FMT in the liver. d The functional enrichment analysis of STZ increased genes while these were decreased by A10-FMT or Con-FMT in the liver. e Blood total triglyceride (TG) levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. f Blood total cholesterol (TC) levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. g Blood total antioxidant capability (T-AOC) levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. h Blood total SOD levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. i Blood catalase levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. j Blood glutathione (GSH) levels. The y-axis represents the relative amount (%). The x-axis represents the treatments. k Western blotting analysis of Bax and Bcl-xl in the liver.